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He was born in the Ukraine of Orthodox parents. In 1595 the Union of Brest brought the Ruthenian Church into communion with Catholic Rome while still preserving its own liturgy. The result was a schism within the church itself, with one party wanting to remain Orthodox and in the orbit of Moscow and Constantinople, while the other accepted the Union. Matters were complicated by the presence of the Greek Uniates, a remnant of a century-old attempt at church union (who remain a living church to this day).

Josaphat joined the first monastery of the order of St Basil to be united to the Catholic Church: he was the first person to do so. He was ordained priest and, eventually and reluctantly, appointed bishop of Polotsk in 1617. Although Sigismund III Vasa, King of Poland and Grand Duke of Lithuania, supported the union, the local aristocracy were against it because it threatened their control of ecclesiastical benefices. Plotting with the Orthodox Patriarch of Jerusalem, who visited the Ukraine in 1621, they stirred up trouble and as a result Josaphat was murdered by a mob in 1623 while on a pastoral visit to Vitebsk.

Take courage, my children, call on God: he will deliver you from tyranny, from the hands of your enemies; for I look to the Eternal for your rescue, and joy has come to me from the Holy One at the mercy soon to reach you from your saviour, the Eternal.